Co-infection with other microorganisms can promote the Candida albicans to be invasive. In this study, Escherichia coli and C. albicans were co-isolated from the women with candidiasis symptoms. The in vitro effects of E. coli on C. albicans hypha development, biofilm formation, antibiotic susceptibility, dispersion from the biofilm, expression of Als3, Hwp1, and Tup1 genes, and pathogenesis in Galleria mellonella were investigated. Electron microscopic images revealed that hypha induction was markedly increased in the bacteria-fungi co-culture. Biofilm formation was increased 2.2 fold in the presence of E. coli. The minimum inhibitory concentration of nystatin against Candida was increased from (μg mL⁻¹) 25 to 50 in the dual biofilm. Candida dissemination was increased up to 2.7 fold from the mixed fungi/bacteria biofilm. The expression of ALS3 and HWP1 genes was increased (5.9 and 2.0 fold, respectively) while the TUP1 gene expression was decreased (0.4 fold) when C. albicans was incubated with E. coli. The simultaneous injection of C. albicans and E. coli to the insect larvae increased Galleria mortality up to 40%. This study demonstrated the effects of E. coli to promote fungi virulence factors, which suggest polymicrobial interaction should be considered during treatment of fungal infections.